The long-term goal of this project is to develop technology to produce transgenic baboons and to make this technology available at the Southwest Regional Primate Research Center and elsewhere to provide a powerful genetic, developmental and physiological nonhuman primate model system for biomedical research. Specific applications of this technology relevant to areas of reproductive biology and contraception and genetics of complex diseases are described herein. However, the availability and optimization of this technology will revolutionize experimental approaches in a wide variety of biomedical areas in nonhuman primate research. Standard methodology currently used to produce transgenics in smaller animals such as mice is prohibitively impractical in nonhuman primates. The approach used will be based on a recently developed technique of producing transgenesis by ICSI as first developed in the mouse by Yanagimachi and coworkers, and recently applied to rhesus monkeys by Schatten and coworkers. A method based on pronuclear injection of IVF embryos will be tested. Recent progress in optimizing superovulation ultrasound monitoring of ovarian stimulation, oocyte harvest, production of embryos by IVF and ICSI and introduction of transgenes into baboon embryos using ICSI is summarized. The goal of the research proposed in this R21 application is to further optimize these technologies for use in the baboon, and in so doing to enhance the efficiency of this approach in nonhuman primates in general. The specific aims of this application include optimization of the following methodologies in the baboon: 1) production of embryos in vitro by IVF and ICSI and subsequent culture to cleavage and blastocyst stages, 2) transfer of IVF embryos to female recipients to foster development to term, and 3) the production of transgenic embryos, including analyses of expression and integration of the transgene and demonstration of tissue-specific transgene expression in embryos. These aims are designed to demonstrate the feasibility and to optimize the efficiency of producing transgenic baboon embryos and the potential to carry these to term to yield transgenic offspring.